Raymond L. Rodriguez | |
---|---|
Born | 1947 (age 75–76) |
Title | Professor, Scientist, Inventor, Entrepreneur |
Raymond L. Rodriguez (born 1947) is an American professor of biology, specializing in molecular biology, genomics and biotechnology. His current research interests include diet-genome interactions, plant-made pharmaceuticals and the food/brain axis. Rodriguez is also an inventor, and entrepreneur. [1] [2] [3] His research at the University of California, San Francisco in the 1970s helped lay the foundation for the biotechnology industry. He also holds several issued US patents. He is involved in programs that promote diversity, equity and inclusion for women and underrepresented minorities in science, technology, engineering, and mathematics (STEM) disciplines. [4] [5]
The son of migrant farm workers, Rodriguez was born in 1947 in Fresno, California and raised in San Joaquin and Kerman, California. In 1965, he graduated from Kerman Union High School. After graduating from Fresno City College, he received a Bachelor of Science degree in biology from California State University, Fresno in 1969 [6] and in the following year he entered the PhD program at the University of California, Santa Cruz. Under the supervision of professor Cedric Davern, [7] Rodriguez produced visual autoradiographic evidence for bidirectional replication [8] of the E. coli chromosome. While a UC Santa Cruz graduate student, Rodriguez received a research fellowship from the Ford Foundation in 1973.
After receiving his PhD in 1974, Rodriguez was awarded an A.P. Giannini postdoctoral fellowship to work with professor Herbert W. Boyer in the department of microbiology at the University of California San Francisco Medical Center. [4] In Boyer's laboratory, Rodriguez collaborated with postdoctoral fellow, Francisco Bolivar Zapata (Paco), to construct more efficient and better characterized cloning vectors. [9] Together, they constructed the 4,361 base pair, circular, autonomously replicating, DNA molecule, pBR322, the first general purpose molecular cloning vector approved [10] by the National Institute of Health Guidelines. [11] [12] The abbreviation, “pBR322,” refers to the plasmid “p,” constructed by Bolivar and Rodriguez “BR,” and the last of “322” transformed colonies to be screened for the pBR322 [13] plasmid. The 1977 publication describing the construction of pBR322 [9] has been cited more than 6,000 times. [14] Soon after its approval by the NIH, pBR322 was used to clone and express the first chemically synthesized gene for the human peptide hormone, somatostatin. [15] The following year, researchers at Harvard University used pBR322 to clone and express rat proinsulin. [16] The main components of pBR322 can be found in many other plasmid vectors, particularly the pUC plasmids designed and constructed by professor Joachim Messing.
In 1976 Rodriguez received fellowships from the National Cancer Institute and the UC President's postdoctoral fellowship program to support plasmid vector research and development. [9]
In 1977, Rodriguez joined the faculty of the University of California, Davis, department of molecular and cellular biology (formerly the genetics department). There, he developed specialized promoter-probe cloning vectors [17] [18] to better understand the regulation of bacterial transcription. In 1998 he received the Distinguished Service Award from the UC Davis College of Agricultural and Environmental Sciences [4] and later their Outstanding Faculty Advisor Award (1992) and the Principles of Community Award (2012), from the UC Davis College of Biological Science. [4]
In 1990, as a member of the Physical Mapping Group, Rodriguez gained experience with genomics by participating in the cloning and mapping of the human APOE gene on chromosome 19 at the Lawrence Livermore National Laboratory [19] Shortly thereafter, he created the International Rice Genome Organization, [20] [21] an ad hoc organization of genomics and agriculture experts to develop a strategy for sequencing the rice genome. [22] This strategy was later used by the Japanese Ministry of Agriculture, Forestry and Fisheries. [23] The first draft of the rice genome was released on April 5, 2002. [24]
In January 2003, Rodriguez received funds from the National Institute on Minority Health and Health Disparities to create a Center of Excellence for Nutritional Genomics. [25] [26] The center was a collaborative effort with the Children's Hospital of Oakland Research Institute. [27] Rodriguez served as Center director until 2009.
From 2007 to 2008, he chaired the Committee of Visitors for the National Science Foundation (NSF) Directorate for Biological Sciences, Plant Genome Research, 3-Year Program Review which assessed the impact of plant genome sequencing on plant biology research. [28]
In 2008 Rodriguez was a Distinguished Lecturer for the USDA-ARS Beltsville Center. [29]
In 2009, Rodriguez received an Honorary Doctorate of Science, from Nara Institute of Science and Technology, Nara, Japan [4] [30] [31]
In 2010, Rodriguez, with the help of professor Somen Nandi, [32] formed Global HealthShare Initiative (GHS), [33] [34] an outreach and knowledge dissemination program. [35] As GHS's executive director, [36] and vice president of Humanity Beyond Barriers, [37] he helped organized international health projects [38] in India, [39] Bangladesh, [40] and Rwanda. [41]
In 2012, the Defense Advanced Research Project Agency (DARPA) funded Rodriguez to engineer a plant-made human butyrylcholinesterase (BuChE), an enzyme used to treat the effects of chemical warfare agents, like sarin gas. [42] Using the fermentation of rice cells transformed with the human BuChE gene, the plant-made enzyme was found to be as effective as human-sourced BuChE in neutralizing sarin. [43]
In 2015, he was an invited presenter to the President's Council of Advisors on Science and Technology (PCAST). [44] [4]
In 2016, Rodriguez was elected a Fellow of the American Association for the Advancement of Science. [5]
In 2018, Rodriguez received the Outstanding Alumni award from Fresno State University, College of Science & Mathematics [4] [2] and was appointed a Distinguish Collaborative Research Professor at Osaka University.
In 2019, Rodriguez directed an interdisciplinary research collaboration project involving the University of California, Davis, Osaka University and Kirin Holdings Co, Japan to use plant cell fermentation to produce safe, effective and affordable human growth factors for stem cell cures. [45] [46]
On June 16, 2021, Osaka University awarded him an honorary degree in recognition of his contributions 'in building the relationship between the two universities, promoting educational exchange, and performing educational and research activity'. [47]
As assistant professor in the UC Davis Department of Genetics, Rodriguez published two edited volumes entitled “Promoters: Structure and Function” in 1982 with M.J. Chamberlin [48] and “Vectors: A Survey of Molecular Cloning Vectors,” in 1987 with D.T. Denhardt. [49] During this period, Rodriguez developed the first molecular cloning lab course in the nation for undergraduates and graduate students. The course was accompanied by a laboratory manual entitled “Recombinant DNA Techniques: An Introduction” [50] co-authored with Dr. Robert C. Tait. [51]
In addition to his research and development of plasmid vectors, Rodriguez also developed a research program to understand the physiological and molecular processes of rice (Oryza sativa). Rodriguez investigated the molecular biology of rice gene systems related to seed germination. This research resulted in the cloning and sequencing of the rice alpha-amylase multigene family. [52] [53] One of the outcomes of this research was the use of alpha-amylase gene promoters to express human proteins in transgenic rice cells. [54] [55] These findings resulted in eighteen issued US patents.
As executive director of the Center of Excellence for Nutritional Genomics, Rodriguez coordinated the research activities of over 50 research faculty, physicians, postdoctoral fellows and graduate students to investigate diet-gene interactions. Center researchers published over 200 research publications and two volumes on diet-gene interactions and their relationship to human health and disease. These included, Nutrigenomics: Discovering the Path to Personalized Nutrition [56] with Dr. Jim Kaput and Nutritional Genomics: Impact of Dietary Regulation of Gene Function on Human Disease [57] with professor Wayne Bidlack. [58] In addition to his duties as Center director, Rodriguez maintained a research program to investigate the role of dietary factors capable of promoting epigenetic changes on genes related to cancer risk. [59]
Throughout his career, Rodriguez provided educational and research opportunities for racial/ethnic minorities (REM) and women in STEM. From 1980 to 1994, he provided research experiences for REMs and women from various California State Universities, NIH MBRS [60] /MARC [61] programs, and select HBCUs around the U.S.. From 1990 to 1993, Rodriguez served as Associate Dean in the UC Davis Office of Graduate Studies. He then organized "Professors for the Future," [62] a mentoring program for diverse, high performing graduate students interested in careers in academia. From 2001 to 2002, Rodriguez served as a member and eventually chair of the Advisory Council, for the National Institute for Minority Health and Health Disparities. From 2005 to 2016 Rodriguez served as a member of the Building Interdisciplinary Research Careers in Women's Health [63] (BIRCWH) program at the UC Davis Medical Center. In 2016, he was a member of the UC Davis NIH Postbaccalaureate Research Education Program [64] (PREP) Internal Advisory and Steering Committees. From 2012 to 2017, Rodriguez served as co-principal investigator and member of the NSF ADVANCE [65] / CAMPOS [66] program designed to increase the number of REM women faculty members in STEM departments at UC Davis. In 2013, Rodriguez received funding from Intel Corporation to organize the first Girls Who Code summer immersion course on a university campus. The Girls Who Code program is designed to close the gender gap in the computer sciences. [67] [2]
A bacterial artificial chromosome (BAC) is a DNA construct, based on a functional fertility plasmid, used for transforming and cloning in bacteria, usually E. coli. F-plasmids play a crucial role because they contain partition genes that promote the even distribution of plasmids after bacterial cell division. The bacterial artificial chromosome's usual insert size is 150–350 kbp. A similar cloning vector called a PAC has also been produced from the DNA of P1 bacteriophage.
Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the transcription of the recombinant DNA to messenger RNA (mRNA), the translation of mRNA into polypeptide chains, which are ultimately folded into functional proteins and may be targeted to specific subcellular or extracellular locations.
A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. The cloning vector may be DNA taken from a virus, the cell of a higher organism, or it may be the plasmid of a bacterium. The vector contains features that allow for the convenient insertion of a DNA fragment into the vector or its removal from the vector, for example through the presence of restriction sites. The vector and the foreign DNA may be treated with a restriction enzyme that cuts the DNA, and DNA fragments thus generated contain either blunt ends or overhangs known as sticky ends, and vector DNA and foreign DNA with compatible ends can then be joined by molecular ligation. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
Chinese hamster ovary (CHO) cells are an epithelial cell line derived from the ovary of the Chinese hamster, often used in biological and medical research and commercially in the production of recombinant therapeutic proteins. They have found wide use in studies of genetics, toxicity screening, nutrition and gene expression, particularly to express recombinant proteins. CHO cells are the most commonly used mammalian hosts for industrial production of recombinant protein therapeutics.
An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. Expression vectors are the basic tools in biotechnology for the production of proteins.
Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination that bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.
A DNA construct is an artificially-designed segment of DNA borne on a vector that can be used to incorporate genetic material into a target tissue or cell. A DNA construct contains a DNA insert, called a transgene, delivered via a transformation vector which allows the insert sequence to be replicated and/or expressed in the target cell. This gene can be cloned from a naturally occurring gene, or synthetically constructed. The vector can be delivered using physical, chemical or viral methods. Typically, the vectors used in DNA constructs contain an origin of replication, a multiple cloning site, and a selectable marker. Certain vectors can carry additional regulatory elements based on the expression system involved.
pBR322 is a plasmid and was one of the first widely used E. coli cloning vectors. Created in 1977 in the laboratory of Herbert Boyer at the University of California, San Francisco, it was named after Francisco Bolivar Zapata, the postdoctoral researcher and Raymond L. Rodriguez. The p stands for "plasmid," and BR for "Bolivar" and "Rodriguez."
A plasmid preparation is a method of DNA extraction and purification for plasmid DNA, it is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology. Many methods have been developed to purify plasmid DNA from bacteria. During the purification procedure, the plasmid DNA is often separated from contaminating proteins and genomic DNA.
A genomic library is a collection of overlapping DNA fragments that together make up the total genomic DNA of a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. The fragments are then inserted into the vector using DNA ligase. Next, the vector DNA can be taken up by a host organism - commonly a population of Escherichia coli or yeast - with each cell containing only one vector molecule. Using a host cell to carry the vector allows for easy amplification and retrieval of specific clones from the library for analysis.
The blue–white screen is a screening technique that allows for the rapid and convenient detection of recombinant bacteria in vector-based molecular cloning experiments. This method of screening is usually performed using a suitable bacterial strain, but other organisms such as yeast may also be used. DNA of transformation is ligated into a vector. The vector is then inserted into a competent host cell viable for transformation, which are then grown in the presence of X-gal. Cells transformed with vectors containing recombinant DNA will produce white colonies; cells transformed with non-recombinant plasmids grow into blue colonies.
Gene delivery is the process of introducing foreign genetic material, such as DNA or RNA, into host cells. Gene delivery must reach the genome of the host cell to induce gene expression. Successful gene delivery requires the foreign gene delivery to remain stable within the host cell and can either integrate into the genome or replicate independently of it. This requires foreign DNA to be synthesized as part of a vector, which is designed to enter the desired host cell and deliver the transgene to that cell's genome. Vectors utilized as the method for gene delivery can be divided into two categories, recombinant viruses and synthetic vectors.
In molecular cloning, a vector is any particle used as a vehicle to artificially carry a foreign nucleic sequence – usually DNA – into another cell, where it can be replicated and/or expressed. A vector containing foreign DNA is termed recombinant DNA. The four major types of vectors are plasmids, viral vectors, cosmids, and artificial chromosomes. Of these, the most commonly used vectors are plasmids. Common to all engineered vectors are an origin of replication, a multicloning site, and a selectable marker.
Joachim Wilhelm "Jo" Messing was a German-American biologist who was a professor of molecular biology and the fourth director of the Waksman Institute of Microbiology at Rutgers University.
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation "pUC" is derived from the classical "p" prefix and the abbreviation for the University of California, where early work on the plasmid series had been conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the most widely used vector molecules as the recombinants, or the cells into which foreign DNA has been introduced, can be easily distinguished from the non-recombinants based on color differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is reversed.
Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine.
Genetic engineering is the science of manipulating genetic material of an organism. The first artificial genetic modification accomplished using biotechnology was transgenesis, the process of transferring genes from one organism to another, first accomplished by Herbert Boyer and Stanley Cohen in 1973. It was the result of a series of advancements in techniques that allowed the direct modification of the genome. Important advances included the discovery of restriction enzymes and DNA ligases, the ability to design plasmids and technologies like polymerase chain reaction and sequencing. Transformation of the DNA into a host organism was accomplished with the invention of biolistics, Agrobacterium-mediated recombination and microinjection. The first genetically modified animal was a mouse created in 1974 by Rudolf Jaenisch. In 1976 the technology was commercialised, with the advent of genetically modified bacteria that produced somatostatin, followed by insulin in 1978. In 1983 an antibiotic resistant gene was inserted into tobacco, leading to the first genetically engineered plant. Advances followed that allowed scientists to manipulate and add genes to a variety of different organisms and induce a range of different effects. Plants were first commercialized with virus resistant tobacco released in China in 1992. The first genetically modified food was the Flavr Savr tomato marketed in 1994. By 2010, 29 countries had planted commercialized biotech crops. In 2000 a paper published in Science introduced golden rice, the first food developed with increased nutrient value.
Genetic engineering techniques allow the modification of animal and plant genomes. Techniques have been devised to insert, delete, and modify DNA at multiple levels, ranging from a specific base pair in a specific gene to entire genes. There are a number of steps that are followed before a genetically modified organism (GMO) is created. Genetic engineers must first choose what gene they wish to insert, modify, or delete. The gene must then be isolated and incorporated, along with other genetic elements, into a suitable vector. This vector is then used to insert the gene into the host genome, creating a transgenic or edited organism.
Recombinant DNA (rDNA), or molecular cloning, is the process by which a single gene, or segment of DNA, is isolated and amplified. Recombinant DNA is also known as in vitro recombination. A cloning vector is a DNA molecule that carries foreign DNA into a host cell, where it replicates, producing many copies of itself along with the foreign DNA. There are many types of cloning vectors such as plasmids and phages. In order to carry out recombination between vector and the foreign DNA, it is necessary the vector and DNA to be cloned by digestion, ligase the foreign DNA into the vector with the enzyme DNA ligase. And DNA is inserted by introducing the DNA into bacteria cells by transformation.
Mervyn James Bibb FRS is an Emeritus Fellow at the John Innes Centre, Norwich, UK.